Investigation on the thermotolerance of porcine ear fibroblasts derived from hybrids with different maternal and paternal genetic background

• Main Authors: Yu, Jun-Xian, 余峻賢
• Other Authors: Shen, Perng-Chih
• Format: Others
• Language: zh-TW
• Published: 2018
• Online Access: http://ndltd.ncl.edu.tw/handle/grfg2r

碩士 === 國立屏東科技大學 === 動物科學與畜產系所 === 106 === It has known that since cytoplasm and nucleus genetic composition of intracellular derived from different thermotolerance varieties, which affect the heat resistance of cell. Furthermore, the cytoplasmic inheritance was belong with maternal inheritance. Therefore, in the present study, we will select purebred Meishan (heat-tolerance), purebred Duroc (heat-sensitivity), KHAPS black pigs (K) of crossbreed from Meishan sow and Duroc boar, KD of crossbreed from K sow and D boar, KDD of crossbreed from KD sow and D boar, DK of crossbreed from D sow and K boar, and DKD of crossbreed from DK sow and D boar. In order to explore the difference of thermotolerance of ear fibroblasts derived from various breeds of porcine with cytoplasmic and nuclear inheritance. The identification of genetic sources in cytoplasm results showed that the DNA PCR-RFLP polymorphisms of mitochondrial DNA D-loop region of ear fibroblasts derived from K, KD and KDD porcine were consistent with that of M porcine, while the cytoplasm were all determined derived from M porcine. However, the DNA polymorphisms of mitochondrial DNA D-loop region of ear fibroblasts derived from DK and DKD porcine were consistent with that of D porcine. Therefore, it was determined that cytoplasms were all derived from D porcine. The apoptotic rates of ear fibroblasts from M porcine in heat-shocked 42.0℃ (24 h: 0.70 vs. 1.33%; 48 h: 1.27 vs. 3.66%; 72 h: 1.73 vs. 6.15%) and 43.0℃ (24 h: 0.76 vs. 1.21%; 48 h: 1.42 vs. 3.75%; 72 h: 2.01 vs. 7.53%) were significantly (P < 0.05) lower than those of cells derived from D porcine with various durations (24-72 h), moreover, genetic in the nucleus (KD vs. DK; KDD vs. DKD) under the same conditions, the apoptotic rates of KD (2.37%) and KDD (2.55%) ear fibroblasts of cytoplasmic inheritance derived from M porcine were significantly (P < 0.05) lower than those of DK (4.03%) and DKD (4.10%) cells of cytoplasmic inheritance derived from D porcine after heat shock (42.0℃, 72 h), respectively. In the protein level, the relative abundances of pro-apoptotic proteins inducing AIF, cytochrome c, Bax, caspase-3, caspase-8 and caspase-9 proteins in ear fibroblasts derived from M porcine were significantly (P < 0.05) lower than those of cells derived from D porcine after heat shock (42.0℃, 72 h). However, genetic in the nucleus (KD vs. DK; KDD vs. DKD) under the same conditions, the relative abundances of cytochrome c, Bax, caspase-3, caspase-8 and caspase-9 proteins in KD and KDD ear fibroblasts of cytoplasmic inheritance derived from M porcine were significantly (P < 0.05) lower than those of DK and DKD cells of cytoplasmic inheritance derived from D porcine after heat shock (42.0℃, 72 h), respectively. Therefore, whether the cytoplasmic inheritance was derived from M or D porcine, the relative abundances of caspase-3, caspase-8 and caspase-9 proteins in those of ear fibroblasts were all significantly (P <0.05) increased in the genetic ratio of D porcine in the nucleus increased after heat shock. Moreover, the relative abundances of Bcl-2/Bax protein ratios in KD and KDD ear fibroblasts of cytoplasmic inheritance derived from M porcine were significantly (P < 0.05) higher than those of DK and DKD cells of cytoplasmic inheritance derived from D porcine after heat shock. In contrast, the relative abundances of HSP-70 protein in ear fibroblasts derived from M, KD and KDD were significantly (P < 0.05) higher than those of cells derived from D, DK and DKD after heat shock. In conclusion, the thermotolerance of ear fibroblasts derived from M porcine is indeed better than those of cells derived from D porcine, moreover, genetic in the nucleus under the same conditions, the thermotolerance of crossbreed porcine ear fibroblasts of cytoplasmic inheritance derived from M porcine are also better than those of cytoplasmic inheritance derived from D porcine, however, the heat resistance of those cells will decrease with the genetic ratio of D porcine in the nucleus increased. Therefore, this also implies that although the cytoplasm of thermotolerance porcine can be improved heat resistance from nucleus of thermosensitive porcine, but the nucleus of thermosensitive porcine still have an adverse effect on the heat resistance of the cells.